The role of PKCepsilon in diabetic retinopathy

PKCepsilon 在糖尿病视网膜病变中的作用

• 批准号: 8018061
• 负责人: Christian Rask-Madsen
• 金额: $ 12.79万
• 依托单位: JOSLIN DIABETES CENTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-01 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8018061
• 关键词: 5' -AMP-activated protein kinase; Acute; Animal Model; Biology; Cell Culture Techniques; Cell Proliferation; Cell physiology; Cells; Cellular biology; Complications of Diabetes Mellitus; Data; Development; Diabetes Mellitus; Diabetic Retinopathy; Disease; Down-Regulation; Drug Delivery Systems; Endothelial Cells; Endothelium; Evaluation; Eye; Fellowship; Genetic; Growth Factor; Injection of therapeutic agent; Institutes; K-Series Research Career Programs; Knock-out; Knockout Mice; Knowledge; Laboratories; Mediating; Mediator of activation protein; Mentors; Minor; Modeling; Molecular; Nitric Oxide; Outcome; Pathway interactions; Patients; Physicians; Play; Postdoctoral Fellow; Prevention; Process; Production; Protein Isoforms; Protein Kinase C; Proteins; Rattus; Research; Research Personnel; Retina; Retinal; Retinal Diseases; Retinal Neovascularization; Role; Scientist; Signal Transduction; Signaling Molecule; Small Interfering RNA; Streptozocin; Testing; Training; Transgenic Mice; Up-Regulation; Vascular Endothelial Growth Factor Receptor; Vascular Endothelial Growth Factor Receptor-2; Vascular Endothelial Growth Factors; Vascular Permeabilities; Wild Type Mouse; Work; angiogenesis; diabetic; diabetic rat; human NOS3 protein; inhibitor/antagonist; innovation; macular edema; mouse model; neovascularization; novel; overexpression; prevent; programs; proliferative diabetic retinopathy; receptor; research study

DESCRIPTION (provided by applicant): Project summary. The applicant is a physician-scientist who has completed his post-doctoral fellowship in the laboratory of Dr. George L. King, who will continue to function as a mentor while the applicant expands his training in new fields, including retinal endothelial cell biology and animal models of retinal complications of diabetes. This training will be co-mentored by Dr. Lloyd Paul Aiello at the Beetham Eye Institute at Joslin Diabetes Center. The Career Development Award would allow the applicant to become an independent researcher in the field of retinal disease and help achieve the long-term objective of understanding growth factor actions in diabetic eye complications. In diabetic macular edema and proliferative diabetic retinopathy, upregulation of vascular endothelial growth factor (VEGF) plays a central role. Protein kinase C (PKC) isoforms a, (3, and e are activated in the diabetic retina, but so far there has been a lack of data from animal models with isoform-specific manipulation of PKC function. In preliminary results, downregulation of PKCe with small interfering RNA (siRNA) in endothelial cell culture had a dramatic effect on several pathways of VEGF-stimulated signaling and cell function, including nitric oxide production and cell proliferation, with minor or opposite effects seen after downregulation of PKCa, (3, or 8. Further, intravitreal injection of PKCe siRNA inhibited VEGF-stimulated retinal vascular permeability in rats. Therefore, the central hypothesis of this application is that PKCe isoform has a quantitatively major role in mediating VEGF-stimulated Akt, eNOS, and AMP-activated protein kinase (AMPK) signaling, and is critical for development of macular edema and proliferative retinopathy in diabetes. The specific aims are: i. To characterize mechanisms of activation of PKCe in retinal endothelial cells and cell function regulated by PKCe; 2. To determine the effect of PKCe knockout or overexpression on vascular permeability during VEGF stimulation and in diabetes; 3. the effect of knockout or overexpression of PKCe function on ischemic neovascularization in the retina. Relevance. The proposed research is expected to have a positive impact on the prevention of macular edema and proliferative retinopathy in patients with diabetes because it will establish the usefulness of the signaling molecule PKCe as a key mediator of these processes and a potential drug target for these conditions.

描述（申请人提供）：项目摘要。申请人是一名医师科学家，他在乔治·L·金（George L. King）博士的实验室完成了博士后研究金，他将继续担任导师，同时申请人在新领域进行培训，包括视网膜内皮细胞生物学和糖尿病视网膜复杂性的动物模型。这项培训将由乔斯林糖尿病中心贝瑟姆眼科研究所的劳埃德·保罗·艾埃洛（Lloyd Paul Aiello）博士共同注册。职业发展奖将使申请人成为视网膜疾病领域的独立研究人员，并有助于实现理解糖尿病眼睛并发症生长因子作用的长期目标。在糖尿病性黄斑水肿和增殖性糖尿病性视网膜病中，血管内皮生长因子（VEGF）的上调起着核心作用。 Protein kinase C (PKC) isoforms a, (3, and e are activated in the diabetic retina, but so far there has been a lack of data from animal models with isoform-specific manipulation of PKC function. In preliminary results, downregulation of PKCe with small interfering RNA (siRNA) in endothelial cell culture had a dramatic effect on several pathways of VEGF-stimulated signaling and cell function, including一氧化氧化物的产生和细胞增殖，在PKCA下调后看到的较小或相反的作用（3或8。此外，玻璃体内注射PKCE siRNA抑制了大鼠VEGF刺激的视网膜血管通透性的视网膜血管通透性。 AMP激活的蛋白激酶（AMPK）信号传导，对于糖尿病中的黄斑水肿和增殖性视网膜病至关重要。 2。确定PKCE敲除或过表达对VEGF刺激和糖尿病中血管渗透性的影响； 3。敲除或过表达PKCE功能对视网膜缺血性新血管形成的影响。关联。预计拟议的研究将对糖尿病患者的黄斑水肿和增殖性视网膜病的预防产生积极影响，因为它将确定信号分子PKCE作为这些过程的关键介体的有用性，并且对这些疾病的潜在药物靶标。